CONCLUSIONI E SVILUPPI FUTUR

Le strategie alimentari chemopreventive possono essere un metodo facilmente attuabile per contrastare i danni che le micotossine determinano sia nell’uomo che nell’animale.

Le molecole individuate in questo studio sono presenti in diversi vegetali quali il rosmarino, la salvia (carnosolo, acido carnosico), il tè verde, il vino rosso, la cioccolata fondente (catechine), arance rosse, mirtilli, melograno (cianidina). Tali vegetali possono normalmente entrare a fare parte della nostra dieta, ma anche in quella animale in forma di estratti miscelabili ai mangimi.

Per approfondire le conoscenze attuali sul meccanismo d’azione delle molecole prese in considerazione, sono in progetto nuovi esperimenti. Tali esperimenti studieranno l’effetto dose/tempo di trattamenti con gli antiossidanti C-3-G, EGCG, ECG, CA e Car sull’attività di alcuni enzimi di fase II quali la glutatione- S-transferasi e la glutatione perossidasi, e sulla disponibilità intracellulare di glutatione ridotto, nelle linee cellulari LLC-PK1 e HepG2.

Altri studi sono in progetto per verificare la capacità chemopreventiva di tali molecole nei confronti dei danni indotti da micotossine, in presenza o in assenza di inibitori specifici della glutatione-S-transferasi.

Si intende poi verificare gli effetti chemopreventivi di estratti vegetali contenenti le molecole studiate. L’utilizzo di estratti vegetali anzichè molecole pure determina una diminuzione dei costi. Inoltre negli estratti potrebbe verificarsi un effetto sinergico tra le molecole presenti.

BIBLIOGRAFIA

Abel S, Gelderblom WC. 1998. Oxidative damage and fumonisin B1-induced toxicity in primary rat hepatocytes and rat liver in vivo. Toxicology 131:121-131. Amorini AM, Fazzina G, Lazzarino G, Tavazzi B, Di Piero D, Santucci R, Simibaldi F, Galvano F, Galvano G. 2001. Activity and mechanism of the antioxidant properties of cyanidin-3-O-beta-glucopyranoside. Free Rad. Res.

35:953-966.

Aruoma OI, Halliwell B, Aeschbach R, Loligers J. 1992. Antioxidant and pro- oxidant properties of active rosemary constituents: carnosol and carnosic acid.

Xenobiotica 22:257-268.

Atroshi F, Biese I, Saloniemi H, Ali-Vehmas T, Saari S, Rizzo A, Veijalainen P. 2000. Significance of apoptosis and its relationship to antioxidants after ochratoxin A administration in mice. J. Pharm. Pharmaceut. Sci. 3:281-291. Autrup H, Bradley KA, Shamsuddinm AKM, Wakhisi J, Wasunna A. 1983. Detection of a putative adduct with fluorescence characteristics identical to 2,3- dihdro-2-(7-guanyl)-3-hydroxy aflatoxin B1 in human urine collected in Muranga district, Kenya. Carcinogenesis 4:1193-1195.

Baudrimont I, Sostarci B, Yenot C, Betbeder AM, Dano-Djedje S, Sanni A, Steyn PS, Creppy EE. 2001. Aspartame prevents the karyomegaly induced by ochratoxin A in rat kidney. Arch. Toxicol. 75:176-183.

Belmadani A, Steyn PS, Tramu G, Betbeder AM, Baudrimont I, Creppy EE. 1999. Selective toxicity of ochratoxin A in primary cultures from different brain regions. Arch. Toxicol. 73:108-114.

Bendele A, Carlton W, Krogh P, Lillehoj EB. 1985. Ochratoxin A carcinogenesis in the (C57BL/6J×C3H) F1 mouse. J. Nation. Canc. Inst. 75:733-742.

Bennett JW and Keller NP. 1997. Mycotoxins and their prevention. In: T. Anke, Editor, Fungal Biotechnology, International Thompson Publishing Company, Weinheim. 265-273.

Briggs T and Rauscher W. 1973. An oscillating iodine clock. J. Chem. Educ.

50:496.

Castegnaro M, Chernozemsky IN, Hietanen E, Bartsch H. 1990. Are mycotoxins risk factors for endemic nephropathy and associated urothelial cancers? Archive

für Geschwulstforschung 60:205-303.

Cervellati R, Höner K, Furrow SD, Neddens C, Costa S. 2001. The Briggs- Rauscher reaction as a test to measure the activity of antioxidants. Helv. Chim.

Acta 84:3533-3547.

Cervellati R, Renzulli C, Guerra MC, Speroni E. 2002. Evaluation of antioxidant activity of some natural polyphenolic compounds using the Briggs-Rauscher reaction method. J. Agric. Food Chem. 50:7504-7509.

Cervellati R, Höner K, Furrow SD, Mazzanti F, Costa S. 2004a. An experimental mechanistic investigation of the complexities arising during inhibition of the Briggs-Rauscher reaction by antioxidants. Helv. Chim. Acta 87:133-155.

Cervellati R, Innocenti G, Dall'Acqua S, Costa S, Sartini E. 2004b. Polyphenols from Polygala spp. and their antioxidant activity. Chem. Biodivers. 1:415-425. Cervellati R, Speroni E, Govoni P, Guerra MC, Costa S, Arnold UW, Stuppner H. 2004c. Wulfenia carinthiaca Jacq. antioxidant and pharmacological activities. Z.

Costa S, Utan A, Speroni E, Cervellati R, Piva G, Prandini A, Guerra MC. 2007. Carnosic acid from rosemary extracts: a potential chemoprotective agent against aflatoxin B1. An in vitro study. J. Appl. Toxicol. 27:152.

Creppy EE, Kane A, Dirheimer G, Lafarge-Frayssinet C, Mousset S, Frayssinet C. 1985. Genotoxicity of ochratoxin A in mice: DNA single-strand break evaluation in spleen, liver and kidney. Toxicol. Lett. 28:29-35.

Del Bano MJ, Lorente J, Castillo J, Benavente-Garcia O, del Rio JA, Ortunto A, Quirin KW, Gerard D. 2003. Phenolic diterpenes, flavones, and rosmarinic acid distribution during the development of leaves, flowers, stems, and roots of Rosmarinus officinalis. Antioxidant activity. J. Agric. Food Chem. 51:4247-4253. Eaton DL, Gallagher EP. 1994. Mechanisms of aflatoxin carcinogenesis. Annu.

Rev. Pharmacol. Toxicol. 34:135-172.

Fink-Gremmels J, Jahn A, Blom MJ. 1995. Toxicity and metabolism of ochratoxin A. Nat. Toxins 3:214-220.

Furrow SD, Cervellati R, Amadori G. 2002. New substrates for the oscillating Briggs-Rauscher Reaction. J. Phys. Chem. 106:5841-5850.

Gallagher EP, Kunze KL, Stapleton PL, Eaton DL. 1996. The kinetics of aflatoxin B1 oxidation by human cDNA-expressed and human liver microsomal cytochromes P450 1A2 and 3A4. Toxicol. Appl. Pharmacol. 141:595-606.

Gautier JC, Holzhaeuser D, Markovic J, Gremaud E, Schilter B, Turesky RJ. 2001. Oxidative damage and stress response from ochratoxin A exposure in rats.

Gekle M. and Silbernagl S. 1996. Renal toxicodynamics of ochratoxin A: a pathophysiological approach. Kidney Blood Press. Res. 19:225-235.

Groopman JD, Dematos P, Egner PA, Love-Hunt A, Kensler TW. 1992. Molecular dosimetry of urinary aflatoxin-N7-guanine and serum aflatoxin-albumin adducts predicts chemoprotection by 1,2-dithiole-3-thione in rats. Carcinogenesis

13:101-106.

Groves CE, Morales M, Wright SH. 1998. Peritubular transport of ochratoxin A in rabbit renal proximal tubules. J. Pharmacol. Exp. Ther. 284:943-948.

Hayes JD, Judah DJ, Neal GE, Nguyen T. 1992. Molecular cloning and heterologous expression of a cDNA encoding a mouse glutathione S-transferase Yc subunit possessing high catalytic activity for aflatoxin B1-8,9-epoxide.

Biochem. J. 285:173-180.

Höner K and Cervellati R. 2002. Measurements of the antioxidant capacity of fruits and vegetables using the BR reaction method. Eur. Food Res. Technol.

215:437-442.

Höner K, Cervellati R, Neddens C. 2002. Measurements of the in vitro antioxidant activity of German white wines using a novel method. Eur. Food Res. Technol.

214:356-360.

Hsieh DP, Cullen JM, Ruebner BH. 1984. Comparative hepatocarcinogenicity of aflatoxins B1 and M1 in the rat. Food Chem. Toxicol. 22:1027-1028.

Huang MT, Huang TW, Lee PH, Chung YC, Hu RH, Lee CS. 1994. Expression of tumor necrosis factor and tissue adhesion molecules in the failed renal allograft.Transplant. Proc. 26:2181-2183.

IARC, 1993. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans. Some Naturally Occurring Substances: Food Items and Constituents, Heterocyclic Aromatic Amines and Mycotoxins. International Agency for Research on Cancer, Lyon. 56:489-521.

Kamp HG, Eisenbrand G, Schlatter J, Würth K, Janzowski C. 2005. Ochratoxin A: Induction of (oxidative) DNA damage, cytotoxicity and apoptosis in mammalian cell lines and primary cells. Toxicology 206:413-425.

Kanner J and Lapidot T. 2001. The stomach as a bioreactor: dietary lipid peroxidation in the gastric fluid and the effects of plant-derived antioxidants. Free

Radc. Biol. Med. 31:1388-1395.

Klassen CD. 2002. Xenobiotic Transporters: another protective mechanism for chemicals. Int. J. Toxicol. 21:7-12.

Kljusuric JG, Djakovic S, Kuhak I, Ganic KK, Komes D, Kurtanjek Z. 2005. Acta

Alimentaria 34:483.

Lee JJ, Jin YR, Hong JT, Kim TJ., Chung JH., Yun YP. 2006. Antiplatelet activity of carnosol is mediated by the inhibition of TXA(2) receptor and cytosolic calcium mobilization. Vascul. Pharmacol.

Lee SJ and Surh YJ. 2005. Nrf2 as a novel molecular target for chemoprevention.

Canc. Lett. 224:171-184.

Malaveille C, Brun G, Bartsch H. 1991. Genotoxicity of ochratoxin A and structurally related compounds in Escherichia coli strains: studies on their mode of action. In: M. Castegnaro, R. Ple tina, G Dirheimer, I.N. Chernozemsky and H. Bartsch, Editors, Mycotoxins, Endemic Nephropathy and Urinary Tract Tumors. IARC Scientific Publications no. 115, International Agency for Research on Cancer, Lyon 261:266.

Masuda T, Inaba Y, Maekawa T, Takeda Y, Tamura H, Yamaguchi H. 2002. Recovery mechanism of the antioxidant activity from carnosic acid quinone, an oxidized sage and rosemary antioxidant. J. Agric. Food Chem. 50:5863-5869. Masuda T, Inaba Y, Takeda Y. 2001. Antioxidant mechanism of carnosic acid: structural identification of two oxidation products. J. Agric. Food Chem. 49:5560- 5565.

Meneghini R and Schumacher RI. 1977. Aflatoxin B1, a selective inhibitor of DNA synthesis in mammalian cells. Chem Biol. Interact. 18:267-276.

Monnet-Tschudi F, Sorg O, Honegger P, Zurich MG, Huggett AC, Schilter B. 1997. Effects of the naturally occurring food mycotoxin ochratoxin A on brain cells in culture. Neurotoxicology 18:831-839.

Moon YJ, Wang X, Morris ME. 2006. Dietary flavonoids: Effects on xenobiotic and carcinogen metabolism. Toxicology in Vitro 20:187-210.

Moreno S, Scheyer T, Romano CS, Vpjnov AA. 2006. Antioxidant and antimicrobial activities of rosemary extracts linked to their polyphenol composition. Free Radic. Res. 40:223-231.

Moss MO. 1996. Centenary review. Mycotoxins. Mycol. Res. 100:513-523.

Mosmann T. 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods 65:55- 63.

Ninomiya K, Matsuda H, Shimoda H, Nishida N, Kasajima N, Yoshino T, Morikawa T, Yoshikawa M. 2004. Carnosic acid, a new class of lipid absorption inhibitor from sage. Bioorg. Med. Chem. Lett. 14:1943-1946.

Niranjan AR, Roy SK. 1982. The use of stomach cytase from the water snail Pila globosa in root tip squashes of ferns. Stain Technol. 57:188-190.

Omar RF, Hasinoff BB, Mejilla F, Rahimtula AD. 1990. Mechanism of ochratoxin A-stimulated lipid peroxidation. Biochem. Pharmacol. 40:1183-1191. Omar RF, Rahimtula AD, Bartsch H. 1991. Role of cytochrome P-450 in ochratoxin A-stimulated lipid peroxidation. J. Biochem. Toxicol. 6:203-209. Palli D, Miraglia M, Saieva C, Masala G, Cava E, Colatosti M, Corsi AM, Russo A, Brera C. 1999. Serum levels of OTA in healthy adults in Tuscany: correlation with individual characteristics and between repeat measurements. Cancer Epidemiology. Biomarkers and Prevention 8:265-269.

Pietta PG. 2000. Flavonoids as antioxidants. J. Nat. Prod. 63:1035-1042. Pitt JI. 2000. Toxigenic fungi and mycotoxins. Br. Med. Bull. 56:184-192.

Pool-Zobel B, Veeriah S, Böhmer FD. 2005. Modulation of xenobiotic metabolising enzymes by anticarcinogens-focus on glutathione S-transferases and their role as targets of dietary chemoprevention in colorectal carcinogenesis. Mut.

Res. 591:74-92.

Prenesti E, Toso S, Berto S. 2005. Redox chemistry of red wine. Quantification by an oscillating reaction of the overall antioxidant power as a function of the temperature. J. Agric. Food Chem. 53:4220-4227.

Ramos-Gomez M, Kwak MK, Dolan PM, Itoh K, Yamamoto M, Tala-lay P, Kensler TW. 2001. Sensitivity to carcinogenesis is increased and chemoprotective efficacy of enzyme inducers is lost in nrf2 transcription factor-deficient mice.

Rahimtula AD, Bereziat JC, Bussacchini-Griot V, Bartsch H. 1998. Lipid peroxidation as a possible cause of ochratoxin A toxicity. Biochem. Pharmacol.

37:4469-4477.

Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic. Biol. Med. 26:1231-1237.

Ringot D, Chango A, Schneider YJ, Larondelle Y. 2006. Toxicokinetics and toxicodynamics of ochratoxin A, an update. Chem. Biol. Interact. 159:18-46. Rizzo AF, Atroshi F, Ahotupa M, Sankari S, Elovaara E. 1994. Protective effect of antioxidants against free radical-mediated lipid peroxidation induced by DON or T-2 toxin. J. Vet. Med. Assoc. 41:81-90.

Schaaf GJ, Nijmeijer SM, Maas RFM, Roestenberg P, Groene EM, Fink- Gremmels J. 2002. The role of oxidative stress in the ochratoxin A-mediated toxicity in proximal tubular cells. Biochim. Biophys. Acta 1588:149-158.

Schwerdt G, Bauer K, Gekle M, Silbernagl S. 1996. Accumulation of ochratoxin A in rat kidney in vivo and in cultivated renal epithelial cells in vitro. Toxicology

114:177-185.

Shen HM, Shi CY, Lee HP, Ong CN. 1994. Aflatoxin B1-induced lipid peroxidation in rat liver. Toxicol. Appl. Pharmacol. 127:145-150.

Shen HM, Ong CN, Lee BL, Shi CY. 1995a. Aflatoxin B1-induced 8- hydroxydeoxyguanosine formation in rat hepatic DNA. Carcinogenesis 16:419- 422.

Shen HM, Ong CN, Shi CY. 1995b. Involvement of reactive oxygen species in aflatoxin B1-induced cell injury in cultured rat hepatocytes. Toxicology 99:115- 123.

Shen HM, Shi CY, Shen Y, Ong CN. 1996. Detection of elevated reactive oxygen species level in cultured rat hepatocytes treated with aflatoxin B1. Free Radical

Biol. Med. 21:139-146.

Sheweita SA and Tilmisany AK. 2003. Cancer and phase II drug-metabolizing enzymes. Curr. Drug Metab. 4:45-58.

Shiraishi F, Curtis LM, Truong L, Poss K, Visner GA, Madsen K, Nick HS, Agarwal A. 2000. Heme oxygenase-1 gene ablation or expression modulates cisplatin-induced renal tubular apoptosis. Am. J. Physiol. Renal Physiol. 278:726- 736.

Speroni E, Cervellati R, Govoni P, Guizzardi S, Renzulli C, Guerra MC. 2003. Efficacy of different Cynara scolymus preparation on liver complaints. J.

Ethnopharmacol 86:203-211.

Sokol DL, Zhang X, Lu P, Gewirtz AM. 1998. Real time detection of DNA/RNA hybridization in living cells. Proc. Natl. Acad. Sci. USA. 95:11538-11543.

Sostaric B and Vukelic M. 1991. Charakterisitcs of urinary tract tumors in the area of Balkan endemic nephropathy in Croatia. In: M. Castegnaro, R. Ple tina, G. Dirheimer, I.N. Chernozemsky and H. Bartsch, Editors, Mycotoxins, Endemic Nephropathy and Urinary Tract Tumors. IARC Scientific Publications no. 115, International Agency for Research on Cancer, Lyon. 29-65.

Sweeney MJ and Dobson ADW. 1998. Review: mycotoxin production by

Aspergillus, Fusarium and Penicillium species. Int. J. Food Microbiol. 43:141-

Talavera S, Felgines C, Texier O, Besson C, Lamaison JL, Remsey C. 2003. Anthocyanins are efficiently absorbed fom the stomach in anestetized rats. J.

Nutr. 133:4178.

Vinson JA, Dabbagh YA, Serry MM, Jang J. 1995. Plant flavonoids, especially tea flavonols, are powerful antioxidant using an in vitro oxidation model for heart disease. J. Agric. Food Chem. 43:5677-5684.

Vukelic M, Sostaric B, Belicza M. 1992. Pathomorphology of Balkan endemic nephropathy. Food Chem. Toxicol. 30:193-200.

Wang SY and Stretch AW. 2001. Antioxidant capacity in cranberry is influenced by cultivar and storage temperature. J. Agric. Food Chem. 49:969-974.

Yin JJ, Smith MJ, Eppley RM, Page SW, Sphon JA. 1998. Effects of fumonisin B1 on lipid peroxidation in membranes. Biochim. Biophys. Acta 1371:134-142. Xu C, Li CY, Kong AN. 2005. Induction of phase I, II and III drug metabolism/transport by xenobiotics. Arch. Pharm. Res. 28:249-268.